JP7427032B2 - Anti-influenza virus composition, respiratory disease treatment composition and anti-aging composition containing black ginseng extract - Google Patents

Description

The present invention relates to an anti-influenza virus composition, a composition for treating respiratory diseases, and an anti-aging composition containing black ginseng extract.

Influenza viruses are classified into four types, such as type A, type B, type C, and type D. Type C only infects humans but is not contagious, and type D does not cause illness in humans. Types A and B are strong viruses that mainly occur in winter and can be transmitted through the human respiratory tract and cause systemic symptoms.

Among these, the serotypes of influenza A viruses are classified according to the types of two proteins on the virus surface, hemagglutinin (H) and neuraminidase (N), and there are 18 types of hemagglutinin and 18 types of neuraminidase (Neuraminidase: N) There are 11 types.

Influenza viruses mutate periodically, making targeted drug treatment difficult. Therefore, neuraminidase inhibitors zanamivir and oseltamivir, which prevent virus proliferation, have been developed and used as drugs. However, recently, oseltamivir has the side effect of inducing suicide.

Therefore, the development of safe natural products while increasing human immunity could be an important means of reducing mortality during influenza pandemics.

Ginseng is a perennial herb belonging to the family Araliaceae and the genus Panax, and is one of the herbal ingredients used in Korean medicine. General physiologically active effects of ginseng are reported to include effects on the central nervous system and anticancer effects (Patent Document 1). In particular, about 40 types of ginsenosides have been discovered so far, and they have a wide range of effects on the endocrine system, metabolic system, etc., including the central nervous system, and are excellent in regulating body functions, that is, normalizing physiological functions. It has been confirmed that it has the same effect. These ginsenosides have similar or opposite effects to each other, but it is known that specific components exert various effects either alone or through interactions of various types.

Among processed products derived from carrots, the present inventor conducted research on raw materials that can improve diseases caused by influenza viruses, and by adjusting the conditions for processing carrots, the content of ginsenosides, which are present in trace amounts in carrots, was reduced. The extract from this black ginseng has been shown to further improve the symptoms of infection caused by influenza virus compared to red ginseng, has excellent efficacy in treating respiratory diseases, and has anti-aging activity. The present invention was completed after confirming that it was very superior.

Korean Published Patent No. 10-2010-0134463

The purpose of this application is to provide a natural substance with excellent anti-influenza virus effects.

The purpose of this application is to provide a natural substance that is highly effective in treating respiratory diseases.

The present application aims to provide a natural substance with excellent anti-aging activity.

As one aspect of achieving the objectives of the present application, one aspect of the present application provides a method for producing black ginseng extract.

Another aspect of the present application provides a composition for preventing, suppressing, or treating diseases caused by influenza viruses.

The composition for preventing, suppressing, or treating diseases caused by influenza viruses of the present application contains black ginseng extract as an active ingredient.

Still other aspects of the present application provide compositions for preventing, suppressing, or treating respiratory diseases.

Still other aspects of the present application provide anti-aging compositions.

The present application will be described in detail below.
An embodiment of the present application includes the steps of steaming ginseng to produce black ginseng; extracting the produced black ginseng using a solvent; and ripening the extracted black ginseng extract. A method for producing a black ginseng extract enriched with ginsenosides Rk1 and Rg5 is provided.

The black ginseng extract is derived from black ginseng obtained by steaming ginseng multiple times.
The ginseng includes Panax ginseng, P. quiquefolius, P. notoginseng, P. japonicus, P. trifolium, and P. ginseng. pseudoginseng), Vietnamese ginseng (P. vietnamensis), or American ginseng (Panax quinquefolium). Ginseng is generally classified into white ginseng and red ginseng depending on the processing method. It is made by steaming and drying ginseng, and various chemical changes occur during the manufacturing process, such as modification of saponins and changes in amino acids. In red ginseng, the heat applied during the manufacturing process produces ginsenoside components that do not exist in ginseng, and the unique active ingredients of red ginseng have the effect of preventing cancer, inhibiting cancer cell growth, lowering blood pressure, and protecting brain nerve cells. , shows an improvement effect on learning ability.

The black ginseng of the present application includes ginseng leaves, young shoots, stems, stem skins, roots, root skins, seeds, fruits, immature fruits, ripe fruits, pulp, pericarp, flowers, androecium, and pistils. (gynoecium), calyx, stamens, petals, calyx lobes, carpels, and combinations thereof can be obtained by steaming nine times. The stamen group refers to all the stamens in one flower, and the pistil group refers to all the pistils in one flower. The carpel is a component that forms the pistil of a flower, and exhibits a modified form of leaves that are generally referred to as flower leaves. The ginseng may be the root, but is not limited thereto, and by mixing other parts of the ginseng that have been steamed nine times, it can be used as an anti-influenza virus.

（前記式で、Ｇｌｃは、グリコシル残基を示す。） The ginsenoside Rk1 is represented by the following formula:

(In the above formula, Glc represents a glycosyl residue.)

（前記式で、Ｇｌｃは、グリコシル残基を示す。） The ginsenoside Rg5 is represented by the following formula:

(In the above formula, Glc represents a glycosyl residue.)

The steaming may be performed at 70° C. to 120° C. 3 to 12 times.

The steaming may be performed at 75°C to 115°C, 80°C to 110°C, 85°C to 105°C, 87°C to 103°C, or 90°C to 100°C. When steaming is performed under the above-mentioned temperature conditions, the content of ginsenosides Rk1 and Rg5, which are active ingredients, is increased, and substances harmful to health are not produced, so that the quality can be optimized.

The steaming may be performed 3 to 11 times, 4 to 10 times, 5 to 9 times, 6 to 9 times, 7 to 9 times, 8 times, or 9 times. When steaming is performed under the above-mentioned steaming conditions, the content of ginsenosides Rk1 and Rg5, which are active ingredients, is increased, and substances harmful to health are not produced, so that the quality can be optimized. As long as the black ginseng extract obtained by applying the steaming conditions of the present application exhibits anti-influenza virus activity against influenza virus infection, the number of times of steaming can be varied.

Steaming is carried out for 1.5 hours to 6 hours, 2 hours or more, 2 hours to 6 hours, 2 hours to 5.5 hours, 2 hours to 5 hours, 2 hours to 4.5 hours, 2 hours per steaming. It may be carried out for hours to 4 hours, 2 hours to 3.5 hours, or 2 hours to 3 hours. When steaming is performed under the above-mentioned time conditions, the content of ginsenosides Rk1 and Rg5, which are active ingredients, is increased, and substances harmful to health are not produced, so that the quality can be optimized.

The method for producing black ginseng extract may further include the step of drying the steamed ginseng every time the steaming is completed. Drying may be performed at 25°C to 60°C, 27°C to 58°C, or 30°C to 55°C. Further, drying may be performed for 10 hours to 30 hours, 12 hours to 28 hours, 12 hours to 26 hours, or 12 hours to 24 hours.

The term "extract" as used in this application means a substance extracted by any method from a raw material, and includes the extract thus extracted, the concentrate obtained thereby, the dried product and powder of the concentrate. It is used as meaning including all without restriction.

The extract is obtained by extracting from a raw material or a dried product thereof, and the raw material for the extract can be cultivated or commercially available, and can be used without restriction.

When obtaining the extract from the raw materials, all known conventional extraction methods can be used, such as solvent extraction, ultrasonic extraction, filtration, and reflux extraction. , can be produced by using a solvent extraction method or a reflux extraction method. The extraction process can be repeated several times, followed by further steps such as concentration or freeze-drying. For example, the obtained extract can be concentrated under reduced pressure to obtain a concentrated solution, and after the concentrated solution is freeze-dried, a highly concentrated extracted powder can be produced using a pulverizer. The extract also includes fractions obtained by additionally fractionating the extract.

The extract may be extracted using water, an organic solvent, or a mixture thereof as an extraction solvent. The organic solvents include alcohol, lower alcohol having 1 to 4 carbon atoms, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, benzene, and these. It may be any one selected from the group consisting of mixed solvents.

When a mixture of water and an organic solvent is used as an extraction solvent, the mixture of water and an organic solvent may be a mixture of water and a lower alcohol having 1 to 4 carbon atoms, or a mixture of water and ethanol; In this case, the solvent is 20% (v/v) to 90% (v/v) ethanol aqueous solution, 25% (v/v) to 85% (v/v) ethanol aqueous solution, 30% (v/v) to 80% (v/v) ethanol aqueous solution, 20% (v/v) to 40% (v/v) ethanol aqueous solution, 30% (v/v) to 60% (v/v) ethanol aqueous solution, 35% (v /v) ~ 75% (v/v) ethanol aqueous solution, 40% (v/v) ~ 70% (v/v) ethanol aqueous solution, 45% (v/v) ~ 65% (v/v) ethanol aqueous solution, Alternatively, it may be a 50% (v/v) to 80% (v/v) ethanol aqueous solution. When producing the black ginseng extract, the extraction solvent may be added in an amount of 4 to 10 times, 5 to 9 times, or 6 to 8 times the amount of black ginseng as an extraction raw material. When extracting under the conditions of the ethanol aqueous solution, the content of ginsenosides Rk1 and Rg5, which are active ingredients, increases.

When preparing the black ginseng extract, the extraction time may be 2 hours to 12 hours, 3 hours to 11 hours, 4 hours to 10 hours, 4 hours to 9 hours, or 4 hours to 8 hours. When extracted under the above time conditions, the content of ginsenosides Rk1 and Rg5, which are active ingredients, increases.

The black ginseng extract may be manufactured through the following steps. After the final steaming is completed, the dried black ginseng is first extracted with a 50% (v/v) to 80% (v/v) ethanol aqueous solution, and the residue generated during the first extraction is extracted with ethanol of the same concentration. Perform a second extraction with an aqueous solution, and perform a tertiary extraction of the residue generated during the second extraction with a 30% (v/v) to 60% (v/v) ethanol aqueous solution. Quaternary extraction can be performed with a 20% (v/v) to 40% (v/v) ethanol aqueous solution. The black ginseng extract may be each extract obtained in each round, or may be a mixture of extracts obtained in the first to fourth extractions. The solvent used for each extraction round can be used in various concentrations. That is, extraction solvents may be used in various combinations to increase the content of ginsenosides Rk1 and Rg5, which are active ingredients.

The black ginseng extract may have a concentration of 55Brix% to 85Brix%, 57Brix% to 83Brix%, 60Brix% to 80Brix%, 62Brix% to 78Brix%, or 65Brix% to 75Brix%. In the case of the Brix% concentration range, the quality such as the content of ginsenosides Rk1 and Rg5, which are active ingredients, is optimized, and the viscosity of the black ginseng extract is not very high, so it can be manufactured in a desired dosage form.

The method for producing black ginseng extract may further include the step of ripening the black ginseng extract after the extraction step.

The step of ripening the black ginseng extract may be performed at a temperature of 80°C or higher, 82°C to 95°C, 84°C to 93°C, 86°C to 91°C, or 88°C to 90°C. When ripening under the above temperature conditions, the contents of ginsenosides Rk1 and Rg5 increase.

The steps of ripening the black ginseng extract include 3 hours or more, 3 hours to 48 hours, 3 hours to 44 hours, 3 hours to 40 hours, 3 hours to 36 hours, 3 hours to 32 hours, 3 hours to 28 hours, Or it may be carried out for 3 to 24 hours. When ripening under the above time conditions, the contents of ginsenosides Rk1 and Rg5 increase.

The total content of ginsenosides Rk1 and Rg5 contained in the black ginseng extract is 100% by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). 20 parts by weight - 90 parts by weight, 25 parts by weight - 85 parts by weight, 30 parts by weight - 80 parts by weight, 35 parts by weight - 75 parts by weight, 40 parts by weight - 70 parts by weight, 45 parts by weight - 65 parts by weight parts, or from 50 parts to 60 parts by weight. When the total content of ginsenosides Rk1 and Rg5 is within the above range, the black ginseng extract is highly effective in preventing, suppressing, or treating diseases caused by influenza viruses.

In the method for producing black ginseng extract, no pressure is applied during steaming. The black ginseng extract of the present application has excellent quality in terms of stability, since harmful substances such as benzopyrene are not generated because pressure is not separately applied during steaming.

The black ginseng is produced by steaming ginseng, and by extracting and ripening the produced black ginseng, a black ginseng extract having a high content of ginsenosides Rk1 and Rg5 can be obtained. For example, black ginseng is produced by steam-ripening carrots at 70°C to 120°C for 3 to 12 times, at least 2 hours per steaming, and Water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof is added 4 to 10 times and extracted for 2 to 12 hours to obtain a black ginseng extract. When aged at temperatures above ℃ for 3 hours or more, the obtained black ginseng extract contains ginsenosides Rk1 and Rg5, which are known to be absent from ginseng or contained in trace amounts in red ginseng. The total amount of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s) is contained in an amount of 20 parts by weight to 90 parts by weight based on the total content of 100 parts by weight. , was significantly higher than that of red ginseng extract.

An embodiment of the present application is a composition for preventing, suppressing, or treating a disease caused by influenza virus, which contains a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is , a composition containing ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s) in an amount of 20 to 90 parts by weight based on a total of 100 parts by weight. provide.

The total content of ginsenosides Rk1 and Rg5 is 25 parts by weight based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). It may be 85 parts by weight, 30 parts by weight to 80 parts by weight, 35 parts by weight to 75 parts by weight, 40 parts by weight to 70 parts by weight, 45 parts by weight to 65 parts by weight, or 50 parts by weight to 60 parts by weight. When the total content of ginsenosides Rk1 and Rg5 is within the above range, the black ginseng extract has excellent effects in preventing, suppressing, or treating diseases caused by influenza virus.

The total content of ginsenoside Rk1 and Rg5 contained in the black ginseng extract is 9 mg/g or more, 9 mg/g to 30 mg/g, 9 mg/g to 28 mg/g, 9 mg/g to 26 mg/g, 9 mg/g. ～24mg/g, 9mg/g～22mg/g, 9.1mg/g～20mg/g, 9.1mg/g～18mg/g, 9.2mg/g～16mg/g, 9.2mg/g～14mg /g, or 9.2 mg/g to 12 mg/g. When the total content of ginsenosides Rk1 and Rg5 is within the above range, the black ginseng extract has excellent effects in preventing, suppressing, or treating diseases caused by influenza virus.

In a specific example of the present application, black ginseng extract containing ginsenosides Rk1 and Rg5 at a concentration of 9.29 mg/g was administered to mice at a concentration of 10 mg/Kg/day once a day to detect influenza A virus (H1N1). was inoculated, and black ginseng extract was administered at a concentration of 10 mg/Kg/day once a day for one week after inoculation. As a result, all mice administered with black ginseng extract survived, and the mortality rate was 0%, similar to the positive control group administered with Tamiflu. On the other hand, when the same method was used but red ginseng extract (containing 0.58 mg/g of ginsenosides Rk1 and Rg5) was administered instead of black ginseng extract, the mice died due to viral infection. It showed a mortality rate of 50%.

The effectiveness of preventing, suppressing, or treating diseases caused by influenza virus is determined by whether the mortality rate is low after influenza virus infection, whether it prevents or improves lung tissue damage caused by influenza virus infection, or whether immune cells are affected early after influenza virus infection. Growth factor (GM-CSF) or immune activation factor (IFN-γ) is highly expressed to increase the proliferation and activity of the immune cells in the early stage of infection and kill the virus, or the immunosuppressive factor (IL-γ) is expressed in the late stage of infection. 10) may be highly expressed to normalize the activated immune system.

The black ginseng extract may further include acidic polysaccharides and polyphenols.

The acidic polysaccharide is 1 mg/g or more, 1 mg/g to 20 mg/g, 1 mg/g to 18 mg/g, 1 mg/g to 16 mg/g, 1 mg/g to 14 mg/g, 1 mg/g to 12 mg/g. , 1.5 mg/g to 10 mg/g, 1 mg/g to 8 mg/g, 2 mg/g to 6 mg/g, 1 mg/g to 4 mg/g, or 2.5 mg/g to 4 mg/g. . When the acidic polysaccharide is contained within the above range, the synergistic effect between the acidic polysaccharide and ginsenosides Rk1 and Rg5 contained in the black ginseng extract becomes high, and the effect of preventing, suppressing, or treating diseases caused by influenza virus is excellent. .

The polyphenol is 15 mg/g or more, 15 mg/g to 40 mg/g, 15 mg/g to 38 mg/g, 15 mg/g to 36 mg/g, 16 mg/g to 34 mg/g, 17 mg/g to 32 mg/g, 18 mg /g to 30mg/g, 19mg/g to 28mg/g, 20mg/g to 26mg/g, or 20mg/g to 24mg/g. When the polyphenol is contained within the above range, the synergistic effect between the polyphenol and the ginsenosides Rk1 and Rg5 contained in the black ginseng extract becomes high, and the effect of preventing, suppressing, or treating diseases caused by influenza virus is excellent.

The influenza virus may be an influenza A virus. The influenza A virus may be H1N1, and the H1N1 may be A/California/04/2009 (H1N1).

The composition for preventing, suppressing, or treating diseases caused by influenza virus, which contains the black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application as an active ingredient, may be a food or a pharmaceutical composition.

The diseases caused by the influenza virus may include, without limitation, all diseases and lesions that can be induced by influenza virus infection. For example, cold, influenza, cough, sneezing, runny nose, muscle pain, sore throat, nasal obstruction, laryngitis, sore throat, hoarseness, headache, sinus pain, rhinitis, pharyngitis, bronchitis, asthma, The condition may be one or more selected from the group consisting of fever, difficulty in breathing, general lethargy, and chills.

The term "prophylaxis" as used in this application refers to all actions by which the compositions of this application delay the onset of disease caused by influenza viruses.

The term "inhibition" as used in this application refers to any action by which the compositions of this application reduce the incidence of disease caused by influenza viruses.

The term "treatment" as used in this application refers to any act by which the compositions of this application cause the symptoms of disease caused by influenza viruses to be ameliorated or beneficial.

The term "administration" as used in this application means introducing a given substance into a subject by some suitable method, and it is understood that the composition of this application is capable of reaching its target in vivo. can be administered via common routes. The administration route of the composition of the present application is not particularly limited, but it can be administered orally or parenterally. Administration of the present application may be carried out from 1 to 4 times, 2 to 3 times, or 2 times per day. Also, the administration of the present application may be carried out for a period of 4 weeks or more, 8 weeks or more, 4 weeks to 12 weeks, or 8 weeks to 12 weeks.

The dosage of the black ginseng extract containing ginsenosides Rk1 and Rg5 contained in the composition for preventing, suppressing, or treating diseases caused by influenza virus depends on the condition and weight of the patient, the degree of disease, the form of the drug, and the route of administration. and the period, but may be selected as appropriate depending on the case. For example, the black ginseng extract containing ginsenosides Rk1 and Rg5 may be administered at a daily dose of 0.0001 to 1000 mg/kg, 0.001 to 700 mg/kg, 0.01 to 500 mg/kg, 0.1 to 100 mg/kg, Alternatively, it may be administered at a dose of 1 to 100 mg/kg, and said administration may be applied once a day or in divided doses. The dosage of the black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application may be increased or decreased depending on the administration route, severity of disease, gender, body weight, age, etc. Therefore, the above dosages are not intended to limit the scope of the present application in any way.

The food composition of the present application may be provided in the form of powder, granules, tablets, capsules, syrup or beverage, and the food composition comprises black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application as active ingredients. It may be used with other foods or food additives. The amount of the active ingredient to be mixed may be appropriately determined depending on the intended use thereof, for example, prevention, health, or therapeutic treatment.

The black ginseng extract containing ginsenosides Rk1 and Rg5 can be added to the food composition in an amount of 0.001% to 60% by weight, such as 0.01% to 50% by weight, 0.1% to 45% by weight, 1% by weight. It may be included in a content of ˜40% by weight, or 1% to 20% by weight. If the content of the black ginseng extract in the cosmetic composition is less than 0.001% by weight, the anti-influenza virus activity of the black ginseng extract may not be sufficiently expressed, and if it exceeds 60% by weight. Although the effect of black ginseng extract may be relatively low compared to the input concentration, there is no problem in terms of safety, so it may be used in an amount above the above range.

There are no particular restrictions on the types of foods in this application, such as meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, and other dairy products, including noodles, gums, and ice cream. , various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes.

The pharmaceutical compositions of the present application may further comprise suitable carriers, excipients or diluents commonly used in the manufacture of pharmaceutical compositions.

Carriers, excipients or diluents that can be used in the pharmaceutical compositions of the present application include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Examples include calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

The pharmaceutical composition of the present application can be prepared into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, by conventional methods. It may be used in the form of a dosage form. Since influenza viruses are transmitted through mucous membranes, the pharmaceutical composition of the present application may be provided in the form of an aerosol, powder, gel, ointment, or drop for easy application to mucous membranes. The mucous membrane may be, but is not limited to, nasal mucosa, oral mucosa, respiratory tract mucosa, or eye mucosa.

When preparing a formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. In such solid preparations, the compound may be added to at least one excipient, such as starch, It may be prepared by mixing calcium carbonate, sucrose or lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as Humectants, sweetening agents, flavoring agents, preservatives, and the like may be included.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. As non-aqueous solvents and suspending agents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used. As a base for suppositories, uitepsol, macrogol, tween 61, cocoa butter, lauric butter, glycerogelatin, etc. may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the pharmaceutical composition of the present application. For example, it is prepared by mixing starch, calcium carbonate, sucrose, lactose and gelatin. In addition to simple excipients, lubricants such as magnesium, stearate, and talc may also be used.

Liquid preparations for oral administration include suspensions, internal solutions, emulsions, and syrups, but in addition to commonly used simple diluents such as water and liquid paraffin, various excipients, For example, humectants, sweetening agents, flavoring agents, preservatives, and the like may be included.

Formulations for dermal administration may be dusting powders, emulsions, suspensions, oils, sprays, ointments, cream pastes, gels, foams, or solutions. The pharmaceutical formulations of the present application may be ointments in anhydrous form, suitable for topical use, containing paraffins, especially low viscosity paraffins, which are liquid at body temperature, or containing natural fats or partially synthetic fats as mentioned above. For example, coconut fatty acid triglycerides, hydrogenated oils, such as hydrogenated peanut oil or castor oil, fatty acid partial esters of glycerol, such as glycerol monostearate and distearate, silicones, such as polymethylsiloxane, such as hexamethyldisiloxane. or may contain octamethyltrisiloxane, for example in conjunction with aqueous creams, fatty alcohols to increase the water absorption capacity, and sterols, wool wax, other emulsifiers and/or other additives. .

The dosage of the black ginseng extract containing ginsenosides Rk1 and Rg5 contained in the pharmaceutical composition of the present application is as described above.

The pharmaceutical composition of the present application can be administered to mammals such as rats, mice, livestock, and humans through various routes. Administration may be administered, for example, by oral, rectal, intravenous, intramuscular, subcutaneous, intrabronchial inhalation, intrauterine dural or intracerebroventricular injection.

The composition for preventing, suppressing, or treating diseases caused by influenza viruses of the present application contains, in addition to the black ginseng extract containing ginsenosides Rk1 and Rg5, one active ingredient that improves, alleviates, treats, or prevents diseases caused by influenza viruses. It may contain more than one species.

The composition for preventing, suppressing, or treating diseases caused by influenza viruses of the present application can be used alone or by surgery, hormonal therapy, drug therapy, and biological reactions for ameliorating, alleviating, treating, or preventing diseases caused by influenza viruses. It may be used in conjunction with methods using modifiers.

As yet another aspect of achieving the object of the present application, the present application includes the step of administering to a subject a composition containing a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient; Provided are methods for preventing, suppressing, or treating diseases caused by influenza viruses.

Compositions and administration for preventing, suppressing, or treating diseases caused by influenza viruses are as described above.

The black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application can be administered in an effective amount to an individual in need thereof.

The individual in need of this may be an individual in need of prevention, suppression, or treatment of a disease caused by influenza virus infection.

As used herein, the term "subject" may be an animal, including humans, or an animal excluding humans.

Another embodiment of the present application is a composition for preventing, suppressing, or treating respiratory diseases that includes a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is: Provided is a composition containing ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s) in an amount of 20 to 90 parts by weight based on a total of 100 parts by weight. do.

The total content of ginsenosides Rk1 and Rg5 is 25 parts by weight based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). It may be 85 parts by weight, 30 parts by weight to 80 parts by weight, 35 parts by weight to 75 parts by weight, 40 parts by weight to 70 parts by weight, 45 parts by weight to 65 parts by weight, or 50 parts by weight to 60 parts by weight. When the total content of ginsenosides Rk1 and Rg5 is within the above range, the black ginseng extract has excellent effects in preventing, suppressing, or treating respiratory diseases.

The total content of ginsenoside Rk1 and Rg5 contained in the black ginseng extract is 9 mg/g or more, 9 mg/g to 30 mg/g, 9 mg/g to 28 mg/g, 9 mg/g to 26 mg/g, 9 mg/g. ～24mg/g, 9mg/g～22mg/g, 9.1mg/g～20mg/g, 9.1mg/g～18mg/g, 9.2mg/g～16mg/g, 9.2mg/g～14mg /g, or 9.2 mg/g to 12 mg/g. When the total content of ginsenosides Rk1 and Rg5 is within the above range, the black ginseng extract has excellent effects in preventing, suppressing, or treating respiratory diseases.

The effect of preventing, suppressing, or treating respiratory diseases is to reduce the accumulation of fine or ultrafine dust in the respiratory organs, such as the bronchi, pharynx, larynx, nasal cavity, sinuses, lungs, etc. This may be due to the effect of regulating or reducing the activity of immune cells in the organ.

The black ginseng extract may further include acidic polysaccharides and polyphenols.

The acidic polysaccharide is 1 mg/g or more, 1 mg/g to 20 mg/g, 1 mg/g to 18 mg/g, 1 mg/g to 16 mg/g, 1 mg/g to 14 mg/g, 1 mg/g to 12 mg/g. , 1.5 mg/g to 10 mg/g, 1 mg/g to 8 mg/g, 2 mg/g to 6 mg/g, 1 mg/g to 4 mg/g, or 2.5 mg/g to 4 mg/g. . When the acidic polysaccharide is contained within the above range, the synergistic effect between the acidic polysaccharide and the ginsenosides Rk1 and Rg5 contained in the black ginseng extract becomes high, and the effect of preventing, suppressing, or treating respiratory diseases is excellent.

The polyphenol is 15 mg/g or more, 15 mg/g to 40 mg/g, 15 mg/g to 38 mg/g, 15 mg/g to 36 mg/g, 16 mg/g to 34 mg/g, 17 mg/g to 32 mg/g, 18 mg /g to 30mg/g, 19mg/g to 28mg/g, 20mg/g to 26mg/g, or 20mg/g to 24mg/g. When the polyphenol is contained within the above range, the synergistic effect between the polyphenol and the ginsenosides Rk1 and Rg5 contained in the black ginseng extract becomes high, and the effect of preventing, suppressing, or treating respiratory diseases is excellent.

The composition for preventing, suppressing, or treating respiratory diseases containing the black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient of the present application may be a food or a pharmaceutical composition.

The respiratory disease may include, without limitation, all diseases that occur in the respiratory organs. For example, the disease may be one or more selected from the group consisting of asthma, chronic obstructive pulmonary disease, bronchitis, pharyngitis, laryngitis, rhinitis, sinusitis, and pneumonia.

The respiratory disease may be a disease induced by fine dust or ultrafine dust. Fine dust refers to particulate matter that is so small that it cannot be seen and is suspended in the atmosphere, and includes particulate matter with a diameter of 50 um or less, 45 um or less, 30 um or less, or 10 um or less. Contains substances. Moreover, ultrafine dust means dust smaller in size than fine dust, and means particulate matter with a diameter of 2.5 um or less. The fine dust and ultrafine dust may be composed of ionic components such as nitrates (NO3-), ammonium ions (NH4+), sulfates (SO42-), carbon compounds, metal compounds, and the like.

The contents described herein relate to a composition for preventing, suppressing, or treating a disease caused by influenza virus, which is another embodiment of the present application, and the content described herein is a composition for preventing, suppressing, or treating a respiratory disease. , and common contents of both will not be described redundantly to avoid undue complexity of the specification. For example, the meanings of the terms "prophylaxis," "suppression," "treatment," and "administration" explained in relation to compositions for preventing, suppressing, or treating diseases caused by influenza viruses, and "dosage of black ginseng extract." ", "Food composition", "Pharmaceutical composition", "Method of administering pharmaceutical composition", "Containing active ingredients other than black ginseng extract", "Use of composition alone or in combination with other methods" The contents may be applied to compositions for preventing, suppressing, or treating respiratory diseases.

As yet another aspect of achieving the object of the present application, the present application provides a method for treating respiratory diseases comprising: administering to an individual a composition comprising a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient; Provide methods of prevention, control, or treatment.

Compositions and administration for preventing, suppressing, or treating respiratory diseases are as described above.

The black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application can be administered in an effective amount to an individual in need thereof.

The individual in need thereof may be an individual in need of prevention, suppression, or treatment of a respiratory disease.

The term "individual" may be an animal including humans or an animal excluding humans.

Still another embodiment of the present application is an anti-aging composition containing black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is ginsenoside Rb1, Rb2, Rc , Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s) in an amount of 20 to 90 parts by weight based on a total of 100 parts by weight.

The total content of ginsenosides Rk1 and Rg5 is 25 parts by weight based on 100 parts by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). It may be 85 parts by weight, 30 parts by weight to 80 parts by weight, 35 parts by weight to 75 parts by weight, 40 parts by weight to 70 parts by weight, 45 parts by weight to 65 parts by weight, or 50 parts by weight to 60 parts by weight. When the total content of ginsenosides Rk1 and Rg5 is within the above range, the black ginseng extract has excellent anti-aging effects.

The total content of ginsenoside Rk1 and Rg5 contained in the black ginseng extract is 9 mg/g or more, 9 mg/g to 30 mg/g, 9 mg/g to 28 mg/g, 9 mg/g to 26 mg/g, 9 mg/g. ～24mg/g, 9mg/g～22mg/g, 9.1mg/g～20mg/g, 9.1mg/g～18mg/g, 9.2mg/g～16mg/g, 9.2mg/g～14mg /g, or 9.2 mg/g to 12 mg/g. When the total content of ginsenoside Rk1 and Rg5 is within the above range, the anti-aging effect of black ginseng extract is excellent.

In this application, the anti-aging effect is determined by the aging-related index factors p15INK4b, p16INK4a, p21, p27, p38, p53, CDK1, CDK2, mechanical target of rapamycin (mTOR), sirtuin 1 (SIRT1), or β-galactos. idase(β -gal) staining can be confirmed by measuring gene and protein expression.

The black ginseng extract may further include acidic polysaccharides and polyphenols.

The acidic polysaccharide is 1 mg/g or more, 1 mg/g to 20 mg/g, 1 mg/g to 18 mg/g, 1 mg/g to 16 mg/g, 1 mg/g to 14 mg/g, 1 mg/g to 12 mg/g. , 1.5 mg/g to 10 mg/g, 1 mg/g to 8 mg/g, 2 mg/g to 6 mg/g, 1 mg/g to 4 mg/g, or 2.5 mg/g to 4 mg/g. . When the acidic polysaccharide is contained within the above range, the synergistic effect between the acidic polysaccharide and the ginsenosides Rk1 and Rg5 contained in the black ginseng extract becomes high, resulting in excellent anti-aging effects.

The polyphenol is 15 mg/g or more, 15 mg/g to 40 mg/g, 15 mg/g to 38 mg/g, 15 mg/g to 36 mg/g, 16 mg/g to 34 mg/g, 17 mg/g to 32 mg/g, 18 mg /g to 30mg/g, 19mg/g to 28mg/g, 20mg/g to 26mg/g, or 20mg/g to 24mg/g. When the polyphenol is contained within the above range, the synergistic effect between the polyphenol and the ginsenosides Rk1 and Rg5 contained in the black ginseng extract becomes high, and the anti-aging effect is excellent.

The anti-aging composition containing the black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application as an active ingredient may be a food or a pharmaceutical composition.

The content described herein with respect to compositions for preventing, suppressing, or treating diseases caused by influenza viruses, which is another embodiment of the present application, may be applied to anti-aging compositions, and both common The subject matter will not be described repeatedly in order to avoid unduly complicating the specification. For example, the meanings of the terms "prophylaxis," "suppression," "treatment," and "administration" explained in relation to compositions for preventing, suppressing, or treating diseases caused by influenza viruses, and "dosage of black ginseng extract." ", "Food composition", "Pharmaceutical composition", "Method of administering pharmaceutical composition", "Containing active ingredients other than black ginseng extract", "Use of composition alone or in combination with other methods" can be applied to the anti-aging composition.

As yet another aspect of achieving the object of the present application, the present application provides prevention of aging, comprising: administering to an individual a composition comprising a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient; Provide a method for suppressing or improving.

The term "improvement" as used in this application refers to all actions by which the compositions of this application improve symptoms that occur with aging.

Anti-aging compositions and administration are as described above.

The black ginseng extract containing ginsenosides Rk1 and Rg5 of the present application can be administered in an effective amount to an individual in need thereof.

The individual in need of this may be an individual in need of prevention, suppression, or improvement of aging.

The term "individual" may be an animal including humans or an animal excluding humans.

Compared to red ginseng, the black ginseng extract produced by the method for producing black ginseng extract of the present application reduces the mortality rate of individuals infected with influenza virus, suppresses or improves lung tissue damage, and is effective in the early stage of influenza virus infection. It increases the proliferation and activity of immune cells and normalizes the immune system, which is activated in the late stages of infection, so it is highly effective in improving the symptoms of infection caused by the influenza virus. Therefore, the black ginseng extract of the present application can be usefully used as an active ingredient in various forms of foods and pharmaceutical compositions for the purpose of improving symptoms of infection by influenza virus.

In addition, the black ginseng extract of the present application has excellent effects on preventing, suppressing, or treating respiratory diseases, so it can be used as an active ingredient in foods and pharmaceutical compositions for preventing or treating respiratory diseases. obtain.

In addition, the black ginseng extract of the present application is very effective in suppressing, preventing, or improving aging, so it may be usefully used as an active ingredient in anti-aging foods and pharmaceutical compositions.

However, the effects of the present application are not limited to the effects mentioned above, and other effects not mentioned can be clearly understood by those skilled in the art from the following description.

FIG. 3 is a diagram showing the amount of change in ginsenosides depending on the steaming time during black ginseng production. FIG. 3 is a diagram showing the amount of change in ginsenosides depending on the steaming time during black ginseng production. FIG. 3 is a diagram showing the amount of change in ginsenosides depending on the steaming time during black ginseng production. FIG. 2 is a diagram showing the amount of change in ginsenosides depending on the aging temperature and aging time of black ginseng concentrate. FIG. 2 is a diagram showing the amount of change in ginsenosides depending on the aging temperature and aging time of black ginseng concentrate. FIG. 2 is a diagram showing the amount of change in ginsenosides depending on the aging temperature and aging time of black ginseng concentrate. It is a graph showing the mortality rate after virus infection in the group treated with standardized black ginseng concentrate. FIG. 3 is a diagram showing the degree of damage to lung tissue after virus infection in the group treated with standardized black ginseng concentrate. FIG. 2 is a graph showing the degree of production of immune activation factors after virus infection in a group treated with standardized black ginseng concentrate. FIG. FIG. 2 is a graph showing the degree of production of immune activation factors after virus infection in a group treated with standardized black ginseng concentrate. FIG. FIG. 2 is a graph showing the degree of production of immune activation factors after virus infection in a group treated with standardized black ginseng concentrate. FIG.

Hereinafter, the present invention will be explained in detail using manufacturing examples and experimental examples.

However, the following production examples and experimental examples are only for illustrating the present invention, and the content of the present application is not limited by the following production examples and experimental examples.

Manufacturing example 1. Production of black ginseng 1-1. Production of red ginseng (black ginseng 1 steamed) The roots of 4-year-old water ginseng were placed in a screw washer (Sangaku FMC, Korea) and washed once for 3 minutes to remove soil or foreign matter. The washed ginseng roots were placed in a steamer and firstly steamed using steam at 98° C. for 2 hours excluding preheating time. After cooling the temperature of the steamer to 55° C., the steamed water ginseng roots were taken out from the steamer. After maintaining the internal temperature of the hot air dryer at 55° C., steamed ginseng roots were added and dried for about 18 hours. The dried red ginseng was transferred to a sun drying field (drying facility using natural light) and dried for over a month. Through the above process, 930 kg of red ginseng (1 steaming) was produced.

1-2. Production of nine steamed nine exposed black ginseng 930 kg of red ginseng produced in Production Example 1-1 was steamed under the same steaming and drying conditions as in Production Example 1-1. The steaming and drying steps described above were repeated, and were called 2 to 9 steamings depending on the number of repetitions. As steaming and drying were repeated, the color gradually became closer to black. Through the above process, 853.8 kg of nine steamed nine exposed black ginseng was produced. When producing the black ginseng extract in Production Example 2 or the black ginseng concentrate in Production Example 3 below, nine steamed nine exposed black ginseng was used as a raw material.

Production example 2. Production of black ginseng extract 284.6 kg of red ginseng produced in Production Example 1-1 and 284.6 kg of black ginseng produced in Production Example 1-2 were taken and ground, and then 70 % (v/v) ethanol aqueous solution was added, and primary extraction was performed at 82° C. for 6 hours. The extraction residue generated during the first extraction was subjected to a second extraction under the same conditions as the first extraction. A 50% (v/v) ethanol aqueous solution was added six times the weight of the extraction residue generated during the second extraction, and the third extraction was performed at 85° C. for 6 hours. A 30% (v/v) aqueous ethanol solution was added six times the weight of the extraction residue generated during the third extraction, and the fourth extraction was carried out at 90° C. for 6 hours. Six times the weight of purified water was added to the weight of the extraction residue generated during the fourth extraction, and the fifth extraction was carried out at 95° C. for 6 hours.

Production example 3. Production of black ginseng concentrate The 1st to 4th red ginseng extracts and the 1st to 4th black ginseng extracts obtained in Production Example 2 were each filtered to separate the extract and the extraction residue. Only the extracts separated from each raw material were placed in a concentrator, and concentrated at 55°C for 4 hours using a vacuum concentrator. 231 kg of red ginseng concentrate having a concentration of 70 Brix% was obtained, and 212 kg of a black ginseng concentrate having a concentration of 70 Brix% were obtained. The obtained red ginseng concentrate and black ginseng concentrate were used as samples for the component analysis and influenza A virus experiment described below.

Experimental example 1. Confirmation of changes in ginsenoside content of black ginseng concentrate depending on steaming time. Steaming was carried out for 9 days under the same steaming conditions as in Production Example 1-1, except that the steaming time was changed to 1 hour, 2 hours, and 3 hours. did. The produced red ginseng was extracted and concentrated according to Production Example 2 and Production Example 3. The content of ginsenoside was measured in the same manner as in Experimental Example 3 below.

When the steaming time was set to 1 hour, as shown in FIG. 1a, it was confirmed that ginsenosides Rk1 and Rg5 were not present even after 9 times of steaming. On the other hand, when the steaming time was set to 2 hours, the content of ginsenosides Rk1 and Rg5 was significantly higher than that of the black ginseng concentrate prepared when the steaming time was set to 1 hour (Figure 1b). In addition, when the steaming time was set to 3 hours, the contents of ginsenosides Rk1 and Rg5 increased compared to when the steaming time was set to 2 hours (Figure 1c).

Experimental example 2. Confirmation of changes in ginsenoside content of black ginseng concentrate by aging temperature or aging time The black ginseng concentrate obtained in Production Example 3 was put into a 2 ml vial, and aged in a water bath at 70°C, 80°C, and 90°C for 8 hours each. , the change in ginsenoside content over time was measured. The content of ginsenoside was measured in the same manner as in Experimental Example 3.

As a result, when aged at 70°C, the content of ginsenosides did not increase even after aging for 8 hours (Figure 2a). On the other hand, when aged at 80°C, the content of ginsenosides Rk1 and Rg5 increased over time, and even when aged at 90°C, the content of ginsenosides Rk1 and Rg5 increased over time, as in the case of aging at 80°C.

Furthermore, as can be seen from FIGS. 2b to 2c, the contents of ginsenosides Rk1 and Rg5 changed depending on the aging time. In the case of black ginseng concentrate that was aged at 80° C. or higher for 3 hours or more, the contents of ginsenosides Rk1 and Rg5 increased.

Experimental example 3. Analysis of ginsenoside components in black ginseng concentrate Analysis of ginsenoside components present in the red ginseng concentrate or black ginseng concentrate obtained in Production Example 3 was performed using HPLC. As a sample, a solution obtained by diluting the black ginseng concentrate obtained in Production Example 3 with methanol at a ratio of 1/30 and filtering it with a 0.45 um filter was used. For HPLC, HPLC1260 (DAD) (Agilent, USA) was used. VENUSIL XBPC18 (4.6 MM x 250 MM, 5.0 um) was used as the column, and as the mobile phase, LC grade water was used as the A solvent and acetonitrile was used as the B solvent. Column temperature was maintained at 35°C. Gradient conditions were: B solvent maintained at 30% at a flow rate of 1 ml/min for 0 to 3.8 minutes, B solvent maintained at 30% at a flow rate of 0.5 ml/min for 3.81 to 4.5 minutes; 4. Increase B solvent from 30% to 42% at a flow rate of 1.2 ml/min for 51 min to 6 min, maintain B solvent at 42% at a flow rate of 0.5 ml/min for 7.5 min to 13 min, 18 Increase B solvent to 47.4% at a flow rate of 0.5 ml/min from 18.1 to 25 minutes, increase B solvent to 55% at a flow rate of 1 ml/min from 18.1 to 25 minutes, and 1 ml/min from 30 to 32 minutes. Increase B solvent from 60% to 90% at a flow rate of 1 ml/min from 42 to 50 minutes, increase B solvent from 60% to 90% at a flow rate of 1 ml/min from 50. was maintained at 30%. The results of component analysis are shown below (unit: mg/g).

As a result of analyzing ginsenosides in black ginseng concentrate, Rb1 0.832mg/g, Rf 1.157mg/g, Rg2(S) 1.008mg/g, Rg2(R) 0.265mg/g, Rh1(S) 0 .941mg/g, Rh1(R) 0.416mg/g, Rg6 0.274mg/g, F4 0.777mg/g, Rk3 0.862mg/g, Rh4 1.714mg/g, Rg3(S) 4.122mg /g, Rg3(R) 1.184mg/g, Rk1 4.747mg/g, and Rg5 4.539mg/g.

When the content of ginsenosides contained in black ginseng concentrate and red ginseng concentrate was compared, in the case of Rk1 and Rg5, it increased about 40 times and about 10 times, respectively, compared to the red ginseng concentrate.

Experimental example 4. Analysis of acidic polysaccharide components in black ginseng concentrate The acidic polysaccharide components present in the red ginseng concentrate and black ginseng concentrate obtained in Production Example 3 were measured using a carbazole sulfuric acid colorimetric method. The samples were used by diluting 300 mg of each concentrate obtained in Production Example 3 with 10 ml of distilled water. The diluted solution was boiled at 90° C. for 3 hours, then cooled and centrifuged (3000 rpm, 10 minutes). 1 ml of the supernatant was taken, and 4 ml of ethanol was added thereto to form a white precipitate. After centrifugation (3000 rpm, 10 min) to obtain a white precipitate, the supernatant was removed, 4 ml of distilled water was added to dissolve the white precipitate, and then n-butanol and CHCl were dissolved in ₁ 1 ml of the solution mixed in step 4 was added, stirred, and centrifuged again (3000 rpm, 10 minutes). From this, 4 ml of the supernatant water extract was taken and sonication was performed.

Put 20 ul of the sonicated solution, 80 ul of distilled water, 50 ul of 0.1% carbazole ethanol reagent (made with 0.125 g of carbazole/100 ml of absolute ethanol), and 600 ul of sulfuric acid into a 2 ml tube, stir, and then transfer to a 96-well plate. The absorbance was measured at 530 nm by adding 200 ul each, and the content was calculated using a calibration curve.

In the case of a standard product, galacturonic acid is dissolved in distilled water and prepared at concentrations of 1000, 500, 250, and 125 mg/L. 20 ul of standard solution, 80 ul of distilled water, 50 ul of 0.1% carbazole ethanol reagent (0.125 g of carbazole) /produced with 100 ml of absolute ethanol) and 600 ul of sulfuric acid were placed in a 2 ml tube and stirred, then 200 ul each was placed in a 96-well plate, the absorbance was measured at 530 nm, and a calibration curve was prepared to measure the concentration. The content of acidic polysaccharide is shown below (unit: mg/g).

As shown in Table 2, the black ginseng concentrate contained a larger amount of acidic polysaccharide than the red ginseng concentrate.

Experimental example 5. Analysis of polyphenol components in black ginseng concentrate The content of polyphenols present in the red ginseng concentrate and black ginseng concentrate obtained in Production Example 3 was measured using Microplate Reader (Powerwave XS, BioTek, USA). Place 2 g of sodium carbonate (Sigma 223484, CAS No. 497-19-8) in a 100 ml constant volume flask, and then add distilled water to make a 100 ml constant volume to prepare a 2% sodium carbonate reagent. Folin-Ciocalteu's phenol reagent (Sigma F9252-1L) and distilled water were mixed in a 1:1 ratio to produce 50% Folin-Ciocalteu's phenol reagent, and aluminum was added to prevent light transmission. covered with foil.

The test solution was prepared by diluting the red ginseng concentrate and black ginseng concentrate obtained in Production Example 3 with distilled water at a ratio of 1:1, taking 0.1 ml each, and adding 0.1 ml of 50% folin-thiocal. The tophenol reagent was mixed with 2 ml of 2% sodium carbonate, left in the dark for 30 minutes, and the absorbance was measured at 750 nm. For the standard solution, 0.4 g of gallic acid (Sigma G7384, CAS No. 149-91-7) was placed in a 100 ml fixed volume flask, and then 100 ml of distilled water was added to a fixed volume of 31.25 ppm and 62.5 ppm. , 125 ppm, 250 ppm, and 500 ppm to prepare standard solutions. For the standard solution, the reagents were mixed in the same manner as for the test solution, left in the dark for 30 minutes, and then the absorbance was measured at 750 nm.

After measuring the absorbance, create a calibration curve with the absorbance of the standard solution on the horizontal axis and the concentration of the standard solution on the vertical axis, and use the following formula 1 to prepare the red ginseng concentrate and black ginseng concentrate obtained in Production Example 3. The total polyphenol content of each liquid was determined.
[Formula 1]
Total polyphenol content (mg/ml) = (A×B×C)/D
*A: Total volume of test solution (ml), B: Dilution factor, C: Concentration of total polyphenols in test solution (mg/ml), D: Sample collection amount (ml)

The content of polyphenols is shown below (unit: mg/g).

As shown in Table 3, the black ginseng concentrate contained a larger amount of polyphenols than the red ginseng concentrate.

Summary of Experimental Examples 3 to 5 The black ginseng concentrate produced in Production Example 3 has higher contents of ginsenosides Rk1, Rg5, acidic polysaccharides, and polyphenols than the red ginseng concentrate produced in Production Example 3. Accordingly, the black ginseng concentrate of Production Example 3 was named a standardized black ginseng concentrate. The standardized black ginseng concentrate was used to measure antiviral activity.

Experimental example 6. Measurement of antiviral activity of black ginseng concentrate In order to measure the effectiveness of the standardized black ginseng concentrate prepared in the above production example in suppressing the new influenza virus, an experiment was conducted as follows. Each experimental group consisted of six 6-week-old BALB/c mice (female) similarly purchased from Semtaco Co., Ltd. and used in the experiment.

Red ginseng concentrate or standardized black ginseng concentrate was administered to mice daily at a concentration of 10 mg/Kg/day for 14 days. Mice used as a negative control group and a positive control group, including mice administered with the red ginseng concentrate or standardized black ginseng concentrate, were each given 30 ul of the new influenza virus (A/California/04/2009 (H1N1)). ) was inoculated into the nose. In the case of mice administered red ginseng concentrate or standardized black ginseng concentrate, red ginseng concentrate or standardized black ginseng concentrate was further administered for one week after influenza virus inoculation. The group that was not vaccinated with influenza virus was defined as the normal group. Among mice infected with influenza virus, a group without sample treatment was set as a negative control group, and among mice infected with influenza virus, a group treated with Tamiflu, a new influenza virus drug, was set as a positive control group. All groups were monitored for survival/lethality of mice for 14 days post-infection. In order to investigate the antiviral immune efficacy mechanism of black ginseng, the presence or absence of lung tissue damage was confirmed 5 days after infection, and immune indicator factor (GM-CSF: Granulocyte-macrophage colony-stimulating factor, IFN-γ: Interferon-gamma, IL-10: Interleukin-10) were measured.

6-1. Evaluation of mortality rate after virus infection The mortality rate after virus infection is shown in Figure 3 and Table 4.

As shown in Figure 3 and Table 4, the negative control group showed a 100% mortality rate after virus infection. On the other hand, the positive control group treated with the antiviral drug Tamiflu had a mortality rate of 0%. In the group administered with red ginseng concentrate, a total of three of the six mice used in the experiment died from virus infection, showing a mortality rate of 50%. However, all six mice in the group administered with standardized black ginseng concentrate survived, and the mortality rate was 0%, similar to the positive control group. Therefore, the standardized black ginseng concentrate had a significantly better protective effect against the new influenza virus than the red ginseng concentrate.

6-2. Evaluation of the degree of damage to lung tissue after virus infection The cause of death of mice infected with the new influenza virus was found to be due to the death of mice 5 days after infection, as shown in the lung tissue of the negative control group or the group administered with red ginseng concentrate shown in Figure 4. It refers to the loss of respiratory function due to tissue damage caused by the accumulation of infectious cells in the lung tissue. The degree of lung tissue damage in the group administered with standardized black ginseng concentrate was relatively lower than that in the group administered with red ginseng concentrate. The standardized black ginseng concentrate was shown to be superior to the standard black ginseng concentrate.

6-3. Evaluation of the degree of immune activation factor production after viral infection The effective stage of treatment for new influenza virus is to effectively remove the virus by multiplying the influenza cells in the early stage of infection and producing a large amount of immune activation factors. After infection, the host returns to a normal state by the production of immunosuppressive factors by infectious cells.

As a result of this experiment, there was no change in the production of immune activation factors in the normal group, as shown by the dotted line in the graph. On the other hand, in the standardized black ginseng concentrate administration group, immune cell growth factor (GM-CSF) was highly expressed on the first day of infection, immune activation factor (IFN-γ) was highly expressed on the third day of infection, and 7 days after infection. On day 1, immunosuppressive factor (IL-10) was highly expressed. Therefore, standardized black ginseng concentrate exhibits antiviral effects by increasing the proliferation and activity of influenza cells in the early stage of infection and killing the virus, and then normalizing the heightened immune system in the late stage of infection. It was found that (see FIGS. 5a to 5c). In addition, many of these immune activation factors showed higher activity compared to the group administered with red ginseng concentrate. was found to be higher than that of

This is considered to be because the standardized black ginseng concentrate has higher contents of ginsenosides Rk1 and Rg5, acidic polysaccharide components, and polyphenols than the red ginseng concentrate.

Experimental example 7. Measurement of the effect of black ginseng concentrate on respiratory health In order to measure the effect of the standardized black ginseng concentrate prepared in the above production example on respiratory health, the following experiment was conducted. Each experimental group consisted of 6 male C57BL/6 mice aged 6 to 8 weeks and was used in the experiment.

Red ginseng concentrate or standardized black ginseng concentrate was administered to mice daily at a concentration of 10 or 250 mg/Kg/day for 14 days. 50 ul of ultrafine dust (SRM2975 10 mg/ml) was administered intratracheally every day for 3 days to mice used as a negative control group, including mice administered with the red ginseng concentrate or standardized black ginseng concentrate. In the case of mice administered red ginseng concentrate or standardized black ginseng concentrate, red ginseng concentrate or standardized black ginseng concentrate was further administered for one week from the day of administration of ultrafine dust. The group to which ultrafine dust was not administered was defined as the normal group. All groups were euthanized 4 to 6 days after the initial injection of ultrafine dust, and then the weight of the lungs and the number of immune (inflammatory) cells were measured to confirm the anti-inflammatory effect of black ginseng. The number was measured. To observe the effect of black ginseng on reducing the accumulation of ultrafine dust in the respiratory lungs, a histopathological examination of the lungs was performed. In addition, in order to investigate the respiratory health functional mechanism against ultrafine black ginseng dust, we investigated immune indicator factors (GM-CSF: granulocyte-macrophage colony-stimulating factor, TNF: tumor necrosis factor, IL-1beta: Interleu). kin-1beta , IL-6, IL-2, & IL-10).

Experimental example 8. Measurement of anti-aging function of black ginseng concentrate In order to measure the effect of the standardized black ginseng concentrate prepared in the above production example on the anti-aging function, the following experiment was conducted. Each experimental group consisted of three 18-month-old C57BL/6 mice (male) and was used in the experiment.

Red ginseng concentrate or standardized black ginseng concentrate was administered to mice daily at a concentration of 300 mg/Kg. All groups were euthanized after daily injections for 28 days. In order to confirm the anti-aging effect of black ginseng, we investigated aging-related indicator factors p15INK4b, p16INK4a, p21, p27, p38, p53, CDK1, CDK2, and mechanical target of rapamycin in liver tissue and hepatocytes isolated from liver tissue. (mTOR), sirtuin 1 (SIRT1), and β-galactosidase (β-gal) staining gene and protein expression were measured.

Manufacturing example 4. Food manufacturing 4-1. Production of Flour Foods 0.5 to 5.0 parts by weight of the black ginseng extract of the present invention was added to wheat flour, and the mixture was used to produce bread, cakes, cookies, crackers, and noodles.

4-2. Preparation of Soup and Gravies 0.2 to 5.0 parts by weight of the black ginseng extract of the present invention was added to soups and gravies to prepare processed meat products for health promotion, noodle soups and gravies.

4-3. Production of ground beef 10 parts by weight of the black ginseng extract of the present invention was added to ground beef to produce ground beef for health promotion.

4-4. Production of Dairy Products 5 to 10 parts by weight of the black ginseng extract of the present invention was added to milk, and the milk was used to produce various dairy products such as butter and ice cream.

4-5. Manufacture of Zen food Brown rice, barley, glutinous rice, and barley were pregelatinized and dried using a known method, roasted, and then ground into powder with a particle size of 60 mesh using a grinder.

Black beans, black sesame, and perilla were also steamed and dried using a known method, roasted, and then ground into powder with a particle size of 60 mesh using a grinder.

The black ginseng extract of the present invention was concentrated under reduced pressure using a vacuum concentrator, sprayed, and dried using a hot air dryer, and the resulting dried product was ground to a particle size of 60 mesh using a grinder to obtain a dry powder.

The grains, seeds and nuts prepared above, and the black ginseng extract of the present invention were blended in the following proportions.

Grains (30 parts by weight of brown rice, 15 parts by weight of pigeon wheat, 20 parts by weight of barley), seeds (7 parts by weight of perilla, 8 parts by weight of black soybeans, 7 parts by weight of black sesame), black ginseng extract of the present invention (3 parts by weight) ), Reishi (0.5 parts by weight), Rhihuang (0.5 parts by weight)

4-6. Manufacture of health drinks Additives such as liquid fructose (0.5%), oligosaccharides (2%), sugar (2%), salt (0.5%), water (75%) and the black color of the present invention. After homogeneously blending with 5 g of ginseng extract and instant sterilization, the mixture was packaged in small packaging containers such as glass bottles and PET bottles.

4-7. Production of vegetable juice Vegetable juice was produced by adding 5 g of the black ginseng extract of the present invention to 1,000 ml of tomato or carrot juice.

4-8. Production of Fruit Juice Fruit juice was produced by adding 1 g of the black ginseng extract of the present invention to 1,000 ml of apple or grape juice.

Manufacturing example 5. Production of pharmaceutical composition 5-1. Production of powder 2 g of black ginseng extract of the present invention
1g lactose
The ingredients were mixed and filled into airtight packages to produce a powder.

5-2. Manufacture of tablets 100 mg of black ginseng extract of the present invention
Corn starch 100mg
Lactose 100mg
Magnesium stearate 2mg
After mixing the above components, the mixture was compressed using a conventional tablet manufacturing method to produce a tablet.

5-3. Production of capsules 100 mg of black ginseng extract of the present invention
Corn starch 100mg
Lactose 100mg
Magnesium stearate 2mg
After mixing the ingredients, the mixture was filled into gelatin capsules using a conventional capsule manufacturing method to produce capsules.

5-4. Production of pills 1 g of black ginseng extract of the present invention
Lactose 1.5g
1g glycerin
xylitol 0.5g
After mixing the above components, the product was prepared in a conventional manner so that each round weighed 4 g.

5-5. Production of granules 150 mg of black ginseng extract of the present invention
Soybean extract 50mg
glucose 200mg
Starch 600mg
After mixing the ingredients, 100 mg of 30% ethanol was added and dried at 60 degrees Celsius to form granules, which were then filled into packages.

Claims (11)

steaming ginseng at 70° C. to 120° C. for 3 to 12 times for more than 2 hours each time to produce black ginseng;
Extracting the produced black ginseng using a solvent;
Aging the extracted black ginseng extract at 80°C or higher for 3 hours or more ,
The black ginseng extract further contains acidic polysaccharides and polyphenols,
The black ginseng extract contains ginsenosides in which the total content of ginsenosides Rk1 and Rg5 is 9 mg/g or more, the acidic polysaccharide content is 1 mg/g or more, and the polyphenol content is 15 mg/g or more. A method for producing a black ginseng extract enriched with Rk1 and Rg5. The total content of ginsenosides Rk1 and Rg5 contained in the black ginseng extract is 100% by weight of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). The method for producing a black ginseng extract according to claim 1, wherein the black ginseng extract is contained in an amount of 20 to 90 parts by weight. A composition for preventing, suppressing, or treating diseases caused by influenza virus, which contains a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is ginsenosides Rb1, Rb2, It is contained in the black ginseng extract in an amount of 20 to 90 parts by weight based on 100 parts by weight of the total content of Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). The total content of ginsenoside Rk1 and Rg5 is 9 mg/g or more,
The black ginseng extract further contains acidic polysaccharides and polyphenols,
A composition for preventing, suppressing, or treating a disease caused by influenza virus, wherein the content of the acidic polysaccharide is 1 mg/g or more, and the content of the polyphenol is 15 mg/g or more. The composition for preventing, suppressing, or treating a disease caused by influenza virus according to claim 3 , wherein the influenza virus is influenza A virus. The composition for preventing, suppressing, or treating a disease caused by influenza virus according to claim 4 , wherein the influenza A virus is H1N1. The composition for preventing, suppressing, or treating a disease caused by influenza virus according to claim 3 , wherein the composition is a food or a pharmaceutical composition. The disease is caused by influenza virus according to claim 3 , wherein the disease is any one or more selected from the group consisting of cold, influenza, sore throat, nasal obstruction, laryngitis, rhinitis, pharyngitis, bronchitis, and asthma. A composition for preventing, suppressing, or treating a disease. A composition for preventing, suppressing, or treating respiratory diseases containing a black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is ginsenosides Rb1, Rb2, Rc, Ginsenoside Rk1 is contained in the black ginseng extract in an amount of 20 to 90 parts by weight based on a total of 100 parts by weight of the content of Rd, Re, Rg1, Rg3(s), Rk1, Rg5 and Rh1(s). and the total content of Rg5 is 9 mg/g or more,
The black ginseng extract further contains acidic polysaccharides and polyphenols,
A composition for preventing, suppressing, or treating respiratory diseases, wherein the acidic polysaccharide content is 1 mg/g or more, and the polyphenol content is 15 mg/g or more. 9. The respiratory disease according to claim 8 , wherein the respiratory disease is selected from the group consisting of asthma, chronic obstructive pulmonary disease, bronchitis, pharyngitis, laryngitis, rhinitis, sinusitis, and pneumonia. A composition for preventing, suppressing, or treating. The composition for preventing, suppressing, or treating a respiratory disease according to claim 9 , wherein the respiratory disease is induced by fine dust or ultrafine dust. An anti-aging composition containing black ginseng extract containing ginsenosides Rk1 and Rg5 as an active ingredient, wherein the total content of ginsenosides Rk1 and Rg5 is ginsenoside Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3 (s ), Rk1, Rg5 and Rh1(s) in a total amount of 20 to 90 parts by weight per 100 parts by weight, and the total content of ginsenosides Rk1 and Rg5 contained in the black ginseng extract is 9 mg/ g or more,
The black ginseng extract further contains acidic polysaccharides and polyphenols,
The anti-aging composition, wherein the content of the acidic polysaccharide is 1 mg/g or more, and the content of the polyphenol is 15 mg/g or more.

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